This recipe will aim to provide a nutrient-rich environment for Bacillus subtilis fermentation.

Ingredients:

  • 750 ml filtered water
  • 250 ml pure apple juice (no added sugars)
  • 30 g miso paste
  • 20 g ground pumpkin seeds
  • 15 ml fish sauce
  • 5 g sea salt
  • B. subtilis starter culture

Equipment:

  • Large glass jar (1.5-2 liter capacity)
  • pH meter or pH strips
  • Thermometer
  • Cheesecloth or coffee filter
  • Rubber band
  • Blender or food processor
  • Stirring rod or spoon

Instructions:

  1. Sterilize all equipment by boiling for 10 minutes or using a pressure cooker.
  2. In a blender, combine the ground pumpkin seeds with 100 ml of the filtered water. Blend until smooth to create a “pumpkin seed milk.”
  3. In the glass jar, whisk together the miso paste and fish sauce until well combined.
  4. Add the apple juice, pumpkin seed milk, and remaining water to the jar. Mix well.
  5. Add the sea salt and stir until dissolved.
  6. Check the pH of the mixture. Aim for a pH between 6.5-7.5. Adjust if necessary using a small amount of baking soda to increase pH or lemon juice to decrease pH.
  7. Cover the jar with cheesecloth or a coffee filter and secure with a rubber band.
  8. Heat the mixture to 75°C (167°F) for 15 minutes to pasteurize, then cool to 37°C (98.6°F). Note: We’re using a lower temperature to preserve more nutrients from the natural ingredients.
  9. Add the B. subtilis starter culture according to the supplier’s instructions.
  10. Incubate at 35-37°C (95-98.6°F) for 24-48 hours, stirring gently once or twice a day.
  11. The fermentation is complete when you see a cloudy appearance and a slightly sour, umami smell.
  12. Strain the liquid through a fine mesh strainer or cheesecloth, then refrigerate to stop further fermentation.

Notes:

  • This recipe uses natural ingredients which may vary in composition. Monitor the fermentation closely and adjust as needed.
  • The apple juice provides sugars for fermentation, miso and fish sauce provide peptides and minerals, and pumpkin seeds offer additional nutrients.
  • Due to the use of fish sauce, this recipe is not vegetarian. For a vegetarian version, omit fish sauce and increase miso paste to 40g.
  • Always ensure proper sanitation to prevent unwanted microbial growth.
  • This recipe is for experimental purposes only. Do not consume without proper safety testing.
  • Fermentation time may vary. Always ensure proper fermentation before using the product.

This recipe incorporates the natural ingredients you specified while aiming to provide a suitable environment for B. subtilis fermentation. Here are some key points about this modified recipe:

  1. Nutrient sources:
  • Apple juice provides sugars and some vitamins
  • Miso offers proteins, minerals, and umami flavors
  • Fish sauce contributes amino acids and minerals
  • Pumpkin seeds provide additional proteins, fats, and minerals
  1. pH balance: The recipe should naturally be close to the preferred pH for B. subtilis, but it’s important to check and adjust if necessary.
  2. Pasteurization: We’re using a lower temperature to preserve more nutrients from the natural ingredients, but this means extra care must be taken to prevent contamination.
  3. Monitoring: Due to the complex nature of these natural ingredients, it’s especially important to monitor the fermentation closely for any signs of unwanted microbial growth.
  4. Straining: The final straining step helps to remove solid particles from the pumpkin seeds and miso.
  5. Flavor: This fermentation will likely have a more complex flavor profile due to the umami-rich ingredients.

Bacillus subtilis has several potential benefits, both as a probiotic organism and in various applications. Here’s an overview of its key benefits:

  1. Probiotic effects:
  • Supports gut health and digestion
  • May help balance the gut microbiome
  • Potential to reduce inflammation in the digestive system
  1. Immune system support:
  • Can stimulate the production of antibodies
  • May enhance the body’s natural defense mechanisms
  1. Nutrient production:
  • Produces vitamin K2 (menaquinone-7)
  • Can synthesize certain B vitamins
  1. Enzyme production:
  • Produces various enzymes that aid in digestion
  • Enzymes have industrial applications (e.g., detergents, food processing)
  1. Agricultural benefits:
  • Used as a biocontrol agent against plant pathogens
  • Can promote plant growth and stress resistance
  1. Antibiotic production:
  • Produces natural antibiotics like subtilin
  • Potential in developing new antimicrobial compounds
  1. Spore formation:
  • Highly resistant spores allow it to survive harsh conditions
  • Useful for creating stable probiotic products
  1. Bioremediation:
  • Can break down certain pollutants in soil and water
  1. Industrial applications:
  • Used in the production of various chemicals and pharmaceuticals
  • Employed in fermentation processes for food products
  1. Research model:
    • Serves as a model organism for studying bacterial processes
  2. Potential therapeutic applications:
    • Being studied for possible roles in treating various health conditions
    • May have anti-inflammatory and antioxidant properties
  3. Competitive exclusion:
    • Can potentially inhibit the growth of harmful bacteria in various environments